This study aimed to evaluate the effects of acetyl-L-carnitine on follicle survival and growth, stromal cell density and extracellular matrix, as well as on the expression of mRNA for nuclear factor erythroid 2-related factor (NRF2), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and peroxiredoxin 6 (PRDX6) in cultured bovine ovarian cortical tissues. Ovarian fragments (3 × 3 × 1 mm) were cultured for 6 days in α-MEM+ alone or supplemented with 10, 50 or 100 μM acetyl-L-carnitine at 38.5°C with 5% CO2 in humidified air. Before (non-cultured tissues) and after culture, the ovarian fragments were fixed in 4% paraformaldehyde for 12 h for histological analysis or stored at –80ºC for mRNA expression analysis of NRF2, SOD, CAT, PRDX6 and GPX1. The results showed that 100 μM acetyl-L-carnitine increased the percentages of morphologically normal follicles and stromal cell density in cultured ovarian tissues. On the other hand, acetyl-L-carnitine did not influence the percentage of collagen in ovarian tissue nor the expression of mRNAs for NRF2, SOD, CAT, PRDX6 and GPX1. In conclusion, 100 μM acetyl-L-carnitine increased follicle survival and stromal cell density in cultured bovine ovarian tissues but does not influence collagen fibre distribution or the expression of mRNAs for NRF2, SOD, CAT, PRDX6 and GPX1.

This study evaluated the effects of capsaicin (CAP) supplementation on the intake, nutrient digestibility, ruminal fermentation, nitrogen balance, microbial protein synthesis and health traits of bulls managed on pasture. Eight crossbred (Holstein x Zebu) cattle averaging 313 ± 31 kg of body weight (BW) were arranged in a replicated 4 × 4 Latin square design (one square of four bulls, rumen fistulated with 4-inch silicone cannulas and one square of four non-fistulated bulls), which were kept on Pangola grass pasture (Digitaria decumbens). Each experimental period consisted of 21 days, with 7 days for data collection (no washout between periods). Bulls were randomly assigned to the following treatments: CON (control): concentrate supplementation at 0.5% live weight (BW) and no additive (CAPCIN®), or concentrate supplementation at 0.5% BW in association with CAPCIN® (NutriQuest, Campinas, Brazil) fed at the inclusion rates of 150 (CAP150), 300 (CAP300) and 450 (CAP450) mg/animal/day. Digestibilities of dry matter, organic matter and neutral detergent fibre showed quadratic responses to CAP supplementation. Rumen pH linearly increased with CAP supplementation. The numbers of lymphocytes and eosinophils were linearly increased with CAP supplementation. The use of encapsulated pepper in supplements of crossbred (Holstein x Zebu) bulls managed on D. decumbens pasture up to 450 mg/animal/day improves nutrients digestibility and ruminal fermentation and can positively influence the health status of beef cattle managed under tropical conditions.

Sucrose yield in sugarcane is a complex process regulated by both environmental and endogenous factors. However, the metabolic balance driving vegetative growth and sucrose accumulation remains poorly understood. Herein, we carried out a comprehensive assessment of carbohydrate dynamics throughout the crop cycle in two sugarcane varieties varying in biomass production, evaluating the carbon metabolism in both leaves and stalks. Our data revealed that the decline in photosynthetic rates during sugarcane maturation is associated not only to accumulation of sugars in leaves but also due to stomatal and non-stomatal limitations. We found that metabolic processes in leaves and stalks were intrinsically linked. While IACSP94-2094 had higher stalk sucrose concentration than IACSP95-5000, this latter produced more biomass. Compared to IACSP95-5000, IACSP94-2094 showed higher sucrose phosphate synthase (SPS) activity in leaves and stalks, along with lower soluble acid invertase (SAI) activity in leaves during the maximum growth stage. Interestingly, IACSP94-2094 also exhibited higher stalk SPS activity and lower stalk SAI activity than IACSP95-5000 during maturation. High biomass production by IACSP95-5000 was associated with higher sucrose synthase (SuSy) and SAI activity in leaves and higher SuSy and soluble neutral invertase (SNI) activity in stalks when compared to IACSP94-2094 during the maximum growth. Despite the contrasting strategies, both varieties displayed similar total sucrose yield, a balance between sucrose concentration and biomass production. This phenomenon implies the presence of a compensatory mechanism in sugarcane, with high biomass production compensating low sucrose accumulation and vice versa.

This study aims to evaluate the effects of N-acetylcysteine (NAC) on bovine oocyte maturation, mitochondrial activity and transzonal projections (TZP), as well as on the levels of reactive oxygen species (ROS) and messenger RNA (mRNA) for catalase (CAT) superoxide dismutase (SOD), periredoxin-6 (Prdx6), glutathione peroxidase (GPx), growth and differentiation factor-9 (GDF9), histone H1Foo, cyclin B1 (CCNB1) and c-Mos. Bovine cumulus–oocyte complexes (COC) of medium-sized antral follicles (3.0–6.0 mm) were prematured in TCM-199 for 8 h at 38.5°C in 5% CO2. After prematuration in the presence of forskolin and C-type natriuretic peptide, COCs were matured in TCM-199 alone or with 0.1, 0.5 or 2.5 mM NAC. Then, oocytes were classified according to the stage of chromatin. Furthermore, mitochondrial activity and intracellular levels of ROS and TZP were also evaluated. The levels of mRNAs for CAT, SOD, Prdx6, GPx, GDF9, H1Foo, CCNB1 and c-Mos were evaluated using real-time polymerase chain reaction (RT-PCR). The results showed that NAC significantly increased the percentages of oocytes with resumption of meiosis when compared with those oocytes matured in control medium. Oocytes had homogeneous mitochondrial distribution, and those cultured with 0.1 and 0.5 mM NAC had lower levels of ROS when compared with the control. In addition, 0.5 mM NAC reduced TZP and the levels of mRNA for CCNB1. In contrast, NAC did not influence the expression of CAT, GPx, Prdx6, SOD, GDF9, H1Foo, and c-Mos. In conclusion, 0.5 mM NAC reduced the levels of ROS, TZP and mRNA for CCNB1, and improved in vitro resumption of meiosis in oocytes from medium-sized bovine antral follicles.

Maternal nutrition is critical in mammalian development, influencing the epigenetic reprogramming of gametes, embryos, and fetal programming. We evaluated the effects of different levels of sulfur (S) and cobalt (Co) in the maternal diet throughout the pre- and periconceptional periods on the biochemical and reproductive parameters of the donors and the DNA methylome of the progeny in Bos indicus cattle. The low-S/Co group differed from the control with respect to homocysteine, folic acid, B12, insulin growth factor 1, and glucose. The oocyte yield was lower in heifers from the low S/Co group than that in the control heifers. Embryos from the low-S/Co group exhibited 2320 differentially methylated regions (DMRs) across the genome compared with the control embryos. We also characterized candidate DMRs linked to the DNMT1 and DNMT3B genes in the blood and sperm cells of the adult progeny. A DMR located in DNMT1 that was identified in embryos remained differentially methylated in the sperm of the progeny from the low-S/Co group. Therefore, we associated changes in specific compounds in the maternal diet with DNA methylation modifications in the progeny. Our results help to elucidate the impact of maternal nutrition on epigenetic reprogramming in livestock, opening new avenues of research to study the effect of disturbed epigenetic patterns in early life on health and fertility in adulthood. Considering that cattle are physiologically similar to humans with respect to gestational length, our study may serve as a model for studies related to the developmental origin of health and disease in humans.

The aim of this study was to establish a functional freezing–thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen–thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

Composite of PZT (lead zirconate titanate) and PLZT (lead lanthanum zirconate titanate) along an organic, renewable, conductive, and flexible matrix have been developed. The conductive paper was obtained through the reaction between the cellulose extracted from banana stem and functionalized MWCNT. This reaction was made in the presence of hydrogen (from H2SO4), which reduced the size of the cellulose by acid hydrolysis and promotes the interaction between its hydroxyls and the carboxyl groups of nanotubes. Afterward, the PZT particles were synthesized and grown up on the conductive paper by microwaves-assisted hydrothermal synthesis (MHS). The composite material was characterized by structure and morphology. Moreover, gas-sensing in the presence of methanol and electrical resistance as a function of temperature were also investigated. In this way, a simple, low-cost and successful synthesis was reported. Also, a promising flexible was obtained using a chemical transformation process from banana residue as source of cellulose.

The sulfentrazone is regularly applied to sugarcane crop harvest residue forPRE control of weedy species, especially in the dry season during the year,but little is known about how rainfall timing or crop residue mass affectthe release of sulfentrazone into the soil and its subsequent effectiveness.Two experiments were conducted to examine the effects of sugarcane residuemass (5, 10, 15, and 20 t ha−1) and rainfall timing (1, 30, and60 d after herbicide application) on sulfentrazone retention. Rainfallvolumes were simulated at 2.5, 5, 10, 20, 35, 50, and 100 mm. A 20-mmrainfall volume was simulated at 7 and 14 d after the first simulated event.The water passing through the straw was collected after each rainfallsimulation. The concentration of sulfentrazone was measured by liquidchromatography and mass spectrometry. The initial 20 mm of rain released themaximum mass of sulfentrazone from the sugarcane residue. The mass ofsugarcane residue affected the amount of sulfentrazone recovered. The amountof sulfentrazone released from the residue was significantly reduced by thepersistence on the residue surface for long periods before the occurrence ofrain. During periods of low rainfall, recommendations for sulfentrazone ratemust take into account losses that occur when applied over the harvestresidues to design a weed-management plan that does not compromise efficacyand duration of the residual effects.

To date, the spatial distribution pattern and density of Brazil nut trees in logged forest stands is unclear across the Amazon basin. We asked the following questions: (1) What are the densities and spatial distributions of Brazil nut juveniles (10 ≤ dbh < 40 cm) and adults (≥ 40 cm dbh) in three selectively logged Brazil nut concessions (1413 ha sampled) in Madre de Dios, Peru; (2) What is the spatial relationship between adults and juveniles (10 ≤ dbh < 30 cm); and (3) What is the spatial relationship between juveniles (10 ≤ dbh <30 cm) and cut stumps (≥ 10 y)? Spatial analyses were conducted using statistics derived from Ripley's K function. Juveniles were aggregated in all three concessions. Results for adult populations rejected the null hypothesis of a random distribution among trees ≥ 40 cm dbh. We did not find an attraction between juveniles and cut-stump locations, nor between adults and juveniles. The strong peaks of aggregation for juveniles and adult Brazil nuts in this study occurred at long distances (300–900 m), suggesting multiple tree canopy gaps as drivers of spatial distribution patterns, either via natural or anthropogenic sources. Our data contribute to a more thorough understanding of Brazil nut population structure in disturbed forests in south-western Amazonia.

This study investigated the emission of subaquatic noise from recreational tourism motorboats, schooners and a sea-bottom mounted water pump. Analyses demonstrated alterations in several whistle (IF: t = 2.42, P = 0.015; FF: t = −2.22, P = 0.025) and calls patterns (MIF: t = −3.13, P = 0.001; MAF: t = −3.49, P = 0.0005; FD: t = −2.21, P = 0.027; D: t = 2.89, P = 0.004), caused primarily by motorboats. Duration of clicks was also modified (D: t = −3.85, P = 0.0001), mainly by the water pump. The frequency range of all noises (0.43–35.8 kHz) overlaps that used by dolphins (1–48 kHz), causing sound emissions changes, with a considerable increase in number of whistles and a reduction in clicks trains. These changes may be a strategy developed by these dolphins to overcome the noise band. Mitigation measures, such as boating regulations and environmental education for the local community, boaters and tourists are needed to conserve the species. The Guiana dolphin population is apparently already suffering, evidenced by diminished residence time and reduced number of individuals entering the inlet during the presence of pleasure craft.

Rhodnius prolixus is a blood-feeding insect that transmits Trypanosoma cruzi and Trypanosoma rangeli to vertebrate hosts. Rhodnius prolixus is also a classical model in insect physiology, and the recent availability of R. prolixus genome has opened new avenues on triatomine research. Glycogen synthase kinase 3 (GSK-3) is classically described as a key enzyme involved in glycogen metabolism, also acting as a downstream component of the Wnt pathway during embryogenesis. GSK-3 has been shown to be highly conserved among several organisms, mainly in the catalytic domain region. Meanwhile, the role of GSK-3 during R. prolixus embryogenesis or glycogen metabolism has not been investigated. Here we show that chemical inhibition of GSK-3 by alsterpaullone, an ATP-competitive inhibitor of GSK3, does not affect adult survival rate, though it alters oviposition and egg hatching. Specific GSK-3 gene silencing by dsRNA injection in adult females showed a similar phenotype. Furthermore, bright field and 4’−6-diamidino-2-phenylindole (DAPI) staining analysis revealed that ovaries and eggs from dsGSK-3 injected females exhibited specific morphological defects. We also demonstrate that glycogen content was inversely related to activity and transcription levels of GSK-3 during embryogenesis. Lastly, after GSK-3 knockdown, we observed changes in the expression of the Wingless (Wnt) downstream target β-catenin as well as in members of other pathways such as the receptor Notch. Taken together, our results show that GSK-3 regulation is essential for R. prolixus oogenesis and embryogenesis.

The aim of this study was to evaluate the purine levels of lambs experimentally infected with Haemonchus contortus. A total of 12 healthy lambs were divided into two groups, composed of 6 animals each: Group A represented the healthy animals (uninfected), while in Group B the animals were infected with 15 000 larvae of H. contortus. Blood was drawn on days 15, 45 and 75 post-infection (PI) in order to perform the purine analysis (ATP, ADP, AMP, adenosine, inosine, hypoxanthine, xanthine and uric acid) by high pressure liquid chromatography (HPLC) in serum. On day 15 PI a significant (P<0·05) increase in the levels of ATP and inosine was observed in the infected animals, unlike the levels of ADP, adenosine, xanthine and uric acid which were reduced. On day 45 PI a significant (P<0·05) increase in the ATP and xanthine levels in infected animals was observed, contrasting with reduced levels of ADP and uric acid. Finally, on day 75 PI an increase occurred in the levels of ATP, adenosine and hypoxanthine in infected lambs, concomitant with a reduction in the levels of ADP and uric acid (P<0·05). These changes in purine levels may influence the inflammatory process and the pathological events.

Intestinal mucositis is an important toxic side effect of 5-fluorouracil (5-FU) treatment. Saccharomyces boulardii is known to protect from intestinal injury via an effect on the gastrointestinal microbiota. The objective of the present study was to evaluate the effect of S. boulardii on intestinal mucositis induced by 5-FU in a murine model. Mice were divided into saline, saline (control)+5-FU or 5-FU+S. boulardii (16 × 109 colony-forming units/kg) treatment groups, and the jejunum and ileum were removed after killing of mice for the evaluation of histopathology, myeloperoxidase (MPO) activity, and non-protein sulfhydryl group (mainly reduced glutathione; GSH), nitrite and cytokine concentrations. To determine gastric emptying, phenol red was administered orally, mice were killed 20 min after administration, and the absorbance of samples collected from the mice was measured by spectrophotometry. Intestinal permeability was measured by the urinary excretion rate of lactulose and mannitol following oral administration. S. boulardii significantly reversed the histopathological changes in intestinal mucositis induced by 5-FU and reduced the inflammatory parameters: neutrophil infiltration (control 1·73 (sem 0·37) ultrastructural MPO (UMPO)/mg, 5-FU 7·37 (sem 1·77) UMPO/mg and 5-FU+S. boulardii 4·15 (sem 0·73) UMPO/mg); nitrite concentration (control 37·00 (sem 2·39) μm, 5-FU 59·04 (sem 11·41) μm and 5-FU+S. boulardii 37·90 (sem 5·78) μm); GSH concentration (control 477·60 (sem 25·25) μg/mg, 5-FU 270·90 (sem 38·50) μg/mg and 5-FU+S. boulardii 514·00 (sem 38·64) μg/mg). Treatment with S. Boulardii significantly reduced the concentrations of TNF-α and IL-1β by 48·92 and 32·21 % in the jejunum and 38·92 and 61·79 % in the ileum. In addition, S. boulardii decreased the concentrations of chemokine (C–X–C motif) ligand 1 by 5-fold in the jejunum and 3-fold in the ileum. Interestingly, S. boulardii reduced the delay in gastric emptying (control 25·21 (sem 2·55) %, 5-FU 54·91 (sem 3·43) % and 5-FU+S. boulardii 31·38 (sem 2·80) %) and induced the recovery of intestinal permeability (lactulose:mannitol ratio: control 0·52 (sem 0·03), 5-FU 1·38 (sem 0·24) and 5-FU+S. boulardii 0·62 (sem 0·03)). In conclusion, S. boulardii reduces the inflammation and dysfunction of the gastrointestinal tract in intestinal mucositis induced by 5-FU.

The conservation of threatened species poses many challenges but through cooperation and pooling of resources, individuals and organizations can work together to achieve better results. Here we describe our experience, working through a governmental and private alliance, studying one of the most threatened mammals in the Southern Cone. The huemul deer Hippocamelus bisulcus, one of two members of the Hippocamelus genus of South America, is endemic to Argentina and Chile and currently inhabits only a small fraction of its former range. Little is known about the huemul because it generally lives in remote areas with a harsh climate and rugged terrain. Using drive counts and fixed width transects over 5 consecutive years (2004–2008) we estimated density and abundance, and examined population changes and social structure, in three coastal huemul populations in the area of the Témpanos and Bernardo fjords of Bernardo O'Higgins National Park, Chile. Our results suggest that synergistic conservation actions, such as cattle removal and poaching control, can lead to the recovery of threatened huemul. The baseline information obtained from our surveys and the lessons learned through this governmental and private alliance will be useful for future monitoring of the huemul in the Patagonian fjords of Chile.

Since the end of the 1990s, there has been an increasing interest in the application of artificial intelligence (AI) planning techniques to solve real-life problems. In addition to characteristics of academic problems, such as the need to reason about actions, real-life problems require detailed knowledge elicitation, engineering, and management. A systematic design process in which Knowledge and Requirements Engineering tools play a fundamental role is necessary in such applications. One of the main challenges in such design process, and consequently in the study of Knowledge Engineering in AI planning, has been the analysis of requirements and their subsequent transformation into an input-ready model for planners. itSIMPLE is a research project dedicated to the study of a project process to support the design phases of real-life planning models. In this paper, we give an overview of itSIMPLE focusing on the main translation processes among a minimal set of representations: from requirements represented in Unified Modeling Language (UML) to Petri Nets and from UML models to planning domain definition language for problem solving.